Thy1 associates with the cation channel subunit HCN4 in Adult Rat Retina

Gloria J. Partida, Tyler W. Stradleigh, Genki Ogata, Iv Godzdanker, Andrew Ishida

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

PURPOSE. The membrane expression and gene promoter of the glycosylphosphatidylinositol (GPI)-anchored protein Thy1 have been widely used to examine the morphology and distribution of retinal ganglion cells in normal eyes and disease models. However, it is not known how adult mammalian retinal neurons use Thy1. Because Thy1 is not a membrane-spanning protein and, instead, complexes with structural and signaling proteins in other tissues, the aim of this study was to find protein partners of retinal Thy1. METHODS. Coimmunoprecipitation, immunohistochemistry, confocal imaging, and patch-clamp recording were used to test for association of Thy1 and HCN4, a cation channel subunit, in adult rat retina. RESULTS. Hyperpolarization of cells immunopanned by an anti-Thy1 antibody activated HCN channels. Confocal imaging showed that individual somata in the ganglion cell layer bound antibodies against Thy1 and HCN4, that the majority of these bindings colocalized, and that some of the immunopositive cells also bound antibody against a ganglion cell marker (Brn3a). Consistent with these results, Thy1 and HCN4 were coimmunoprecipitated by magnetic beads coated with either anti-Thy1 antibody or anti-HCN4 antibody. In control experiments, beads coated with these antibodies did not immunoprecipitate a photoreceptor rim protein (ABCR) and uncoated beads did not immunoprecipitate either Thy1 or HCN4. CONCLUSIONS. This is the first report that Thy1 colocalizes and coimmunoprecipitates with a membrane-spanning protein in retina, that Thy1 complexes with an ion channel protein in any tissue, and that a GPI-anchored protein associates with an HCN channel subunit protein.

Original languageEnglish (US)
Pages (from-to)1696-1703
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume53
Issue number3
DOIs
StatePublished - Mar 2012

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Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels
Retina
Cations
Anti-Idiotypic Antibodies
Glycosylphosphatidylinositols
Proteins
Ganglia
Antibodies
Membrane Proteins
Retinal Neurons
Eye Diseases
Retinal Ganglion Cells
Protein Subunits
Carisoprodol
Ion Channels
Immunohistochemistry
Gene Expression
Membranes

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Partida, G. J., Stradleigh, T. W., Ogata, G., Godzdanker, I., & Ishida, A. (2012). Thy1 associates with the cation channel subunit HCN4 in Adult Rat Retina. Investigative Ophthalmology and Visual Science, 53(3), 1696-1703. https://doi.org/10.1167/iovs.11-9307

Thy1 associates with the cation channel subunit HCN4 in Adult Rat Retina. / Partida, Gloria J.; Stradleigh, Tyler W.; Ogata, Genki; Godzdanker, Iv; Ishida, Andrew.

In: Investigative Ophthalmology and Visual Science, Vol. 53, No. 3, 03.2012, p. 1696-1703.

Research output: Contribution to journalArticle

Partida, GJ, Stradleigh, TW, Ogata, G, Godzdanker, I & Ishida, A 2012, 'Thy1 associates with the cation channel subunit HCN4 in Adult Rat Retina', Investigative Ophthalmology and Visual Science, vol. 53, no. 3, pp. 1696-1703. https://doi.org/10.1167/iovs.11-9307
Partida, Gloria J. ; Stradleigh, Tyler W. ; Ogata, Genki ; Godzdanker, Iv ; Ishida, Andrew. / Thy1 associates with the cation channel subunit HCN4 in Adult Rat Retina. In: Investigative Ophthalmology and Visual Science. 2012 ; Vol. 53, No. 3. pp. 1696-1703.
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